3.

2 TOOTHPICKASE LAB ACTIVITY Hybrid Mode

Background: Enzymes are proteins that help speed up (catalyze) chemical reactions without being used
up or changed by the reactions. Enzymes are able to increase the rate of chemical reactions by lowering
the activation energy to start the reaction. Each enzyme is specifically designed to fit with a specific
substrate and the enzyme and substrate link at the active site. Various factors impact enzyme activity
including pH, temperature, salinity and concentration of the enzyme and/or substrate.

In this activity, your hand will represent “toothpickase”, an enzyme that aids in breaking apart
toothpicks (the substrate). Your thumb and fingers used to break the toothpick represent the active site
at which the toothpick can be broken. Enzymes work randomly, so to model this you will complete all
tasks with your eyes closed. During this activity, you will examine “normal” toothpickase activity and
the impact of various factors on enzyme activity.

Ground Rules:
 You can use ONLY ONE hand to break the toothpicks in half. You may not use any other part of
your body, and no other objects (table surface, etc.).
 Only ONE toothpick may be broken at a time.
 The toothpick must be broken COMPLETELY in half, or it does not count as a product.
 Each broken toothpick must go back into the original pile (simulating that products and reactants
continue mixing during the reaction).
 Complete all rounds with your eyes CLOSED (simulating the randomness of enzymes)

Materials:
 150 flat toothpicks  Tape
 Stopwatch or timer

Procedure:
Part I: Enzyme Only.

1. Count out 50 toothpicks.

2. Spread the toothpicks out in a small area on the desk/lab table.
3. When the timer says “GO”, the student playing the part of toothpickase will break as many
toothpicks as possible (following the rules above).
4. When the timer says “STOP”, the student playing the part of toothpickase will stop and the
counter will count how many toothpicks were broken. (Remember, the toothpick must be broken
COMPLETELY in half to count as a product.)
5. The data recorder will record the total number of toothpicks broken in Table I.
6. After each round, all toothpicks, broken and unbroken remain for the next round. **If
toothpickase is able to break all toothpicks before the final time interval is finished, record the
time at which the last toothpick was broken.**
a. Round 1: 10 second interval
b. Round 2: 20 second interval (30 seconds total)
c. Round 3: 30 second interval (60 seconds total)
d. Round 4: 60 second interval (120 seconds total)
Part II: Competitive Inhibitor
1. Repeat everything from Part I, but this time start with 30 normal toothpicks and 20 purple
toothpicks. Record data in Table II.

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 Part III: Allosteric Inhibitor
1. Repeat everything from Part I, but this time tape your pinky, ring, and middle finger to your
palm. Leave only thumb and index finger free. Record data in Table III.

Part IV: Allosteric Activator

1. Repeat everything from Part I, but this time one or more teammates can assist, as long as they
don’t break any toothpicks themselves. Record data in Table IV.

Part V: Enzyme Concentration

1. Repeat everything from Part I, but this time multiple group members can break toothpicks. All
rules still apply, eyes closed, etc. Record data in Table V.

Table I: Rate of Toothpickase Activity – Enzyme Only

Round Total # Rate of Activity (#

Interval Cumulativ Time Toothpicks toothpicks
Numbe
Time e Time Interval Broken broken/sec)
r
1 10 10 0-10 3 0.30/sec
2 20 30 10-30 8 0.40/sec

3 30 60 30-60 11 0.36/sec

4 60 120 60-120 15 0.25/sec

Table II: Rate of Toothpickase Activity – Competitive Inhibitor

Round Total # Rate of Activity (#

Interval Cumulativ Time Toothpicks toothpicks
Numbe
Time e Time Interval Broken broken/sec)
r
1 10 10 0-10 6 0.60/sec
2 20 30 10-30 7 0.35/sec

3 30 60 30-60 2 0.07/sec

4 60 120 60-120 10 0.16/sec

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 Table III: Rate of Toothpickase Activity – Allosteric Inhibitor

Round Total # Rate of Activity (#

Interval Cumulativ Time Toothpicks toothpicks
Numbe
Time e Time Interval Broken broken/sec)
r
1 10 10 0-10 2 0.20/sec
2 20 30 10-30 8 0.40/sec

3 30 60 30-60 8 0.26/sec

4 60 120 60-120 5 0.08/sec

Table IV: Rate of Toothpickase Activity – Allosteric Activator

Round Total # Rate of Activity (#

Interval Cumulativ Time Toothpicks toothpicks
Numbe
Time e Time Interval Broken broken/sec)
r
1 10 10 0-10 7 0.70/sec
2 20 30 10-30 13 0.65/sec

3 30 60 30-60 12 0.40/sec

4 25 120 60-120 13 0.52/sec

Table V: Rate of Toothpickase Activity – Increased Enzyme Concentration

Round Total # Rate of Activity (#

Interval Cumulativ Time Toothpicks toothpicks
Numbe
Time e Time Interval Broken broken/sec)
r
1 10 10 0-10 8 0.80/sec
2 20 30 10-30 8 0.40/sec

3 30 60 30-60 12 0.40/sec

4 60 120 60-120 5 0.08/sec

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 Toothpickase Lab
Conclusion Questions:

1. Briefly describe how the five different real-world manipulations were analogies for the actual
effect that those variables have on the rate of enzyme action.
The five real world manipulations represented what happens with enzyme action. Table 1 is a
normal enzyme reaction. Table 2 is when an inhibitor that resembles the normal substrate binds
to the enzyme and prevents the substrate from binding, which is the competitive variable. Table 3
represent allosteric inhibition. When an allosteric inhibitor binds to an enzyme, all active sites on
the protein subunits are changed slightly so that they work less well. This is why the numbers
were slower than other reactions. Table 4 is the same thing but adding a catalyst. Table 5 is
increasing enzyme concentration which speeds up reaction rate. This was represented by adding
team members to break the toothpicks.

2. What happened to the reaction rate as the number of unbroken toothpicks decreased? Why did
this happen?
The reaction rate when down because the subject had to look and find the toothpicks that were
not broken and snap those. There was not a separate pile for the broken and unbroken toothpicks,
they were all scattered about.

3. What happened to the reaction rate when the enzyme concentration was increased? Why?
When the enzyme concentration was increased, the reaction rate was slower. The reaction rate
started off quick but then went down.

4. Name one factor that we did not simulate that can affect the rate of enzyme action.
One of the things that can affect the rate of enzyme action is temperature. Colder temperatures
slow down the rate of reaction while warmer temperatures speed up reactions.

5. What are 3 environmental variables that can affect the rate of enzyme activity?
Temperature, pH enzyme concentration, substrate concentration

6. In 3-4 sentences, summarize what you have learned about enzymes during this activity.
I have learned that there are many different factors that can control the reaction rate. In data table
4, you can see that the numbers were, on average, higher than the numbers in other data tables.
This was because somebody was able to help the person who was breaking the toothpicks. This
is the same this as what a catalyst does to an enzyme reaction.

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